Investigation of an Immunoassay with Broad Specificity to Quinolone Drugs by Genetic Algorithm with Linear Assignment of Hypermolecular Alignment of Data Sets and Advanced Quantitative Structure–Activity Relationship Analysis

Abstract

A polyclonal antibody against the quinolone drug pazufloxacin (PAZ) but with surprisingly broad specificity was raised to simultaneously detect 24 quinolones (QNs). The developed competitive indirect enzyme-linked immunosorbent assay (ciELISA) exhibited limits of detection (LODs) for the 24 QNs ranging from 0.45 to 15.16 ng/mL, below the maximum residue levels (MRLs). To better understand the obtained broad specificity, a genetic algorithm with linear assignment of hypermolecular alignment of data sets (GALAHAD) was used to generate the desired pharmacophore model and superimpose the QNs, and then advanced comparative molecular field analysis (CoMFA) and advanced comparative molecular similarity indices analysis (CoMSIA) models were employed to study the three-dimensional quantitative structure-activity relationship (3D QSAR) between QNs and the antibody. It was found that the QNs could interact with the antibody with different binding poses, and cross-reactivity was mainly positively correlated with the bulky substructure containing electronegative atom at the 7-position, while it was negatively associated with the large bulky substructure at the 1-position of QNs.

Application of quantitative structure‐activity relationship analysis on an antibody and alternariol‐like compounds interaction study

“…The interaction between antigen and antibody of the immunoassay method was influenced by various factors. More and more studies have shown that the specificity of the antibody is of great significance for improving immunoassays . An antibody with high specificity (low cross‐reactivity [CR]) is suitable to be used in the immunoassay detecting a single compound, while an antibody with broad specificity (high CR) is suitable to be used in the immunoassay detecting multiple compounds .…”

“…More and more studies have shown that the specificity of the antibody is of great significance for improving immunoassays. [9][10][11] An antibody with high specificity (low cross-reactivity [CR]) is suitable to be used in the immunoassay detecting a single compound, while an antibody with broad specificity (high CR) is suitable to be used in the immunoassay detecting multiple compounds. [12][13][14][15] However, because of the lack of knowledge about the antibody recognition and the ambiguous structure-activity relationship, artificial control of the specificity of the antibody remains a huge challenge.…”

Ciprofloxacin and Clinafloxacin Antibodies for an Immunoassay of Quinolones: Quantitative Structure–Activity Analysis of Cross-Reactivities

“…Quantitative regulations of immune recognition were recently formulated for monoclonal antibodies against ciprofloxacin (CIP) [23]. A potential of sarafloxacin [29], pazufloxacin [37], marbofloxacin, benfloxacin, norfloxacin, and pefloxacin [37] for broad-specific detection of FQs was also demonstrated. Polyclonal antibodies against clinafloxacin (CLI) were obtained and used for the immunoassay of FQs in milk [38].…”

Substructure-activity relationship studies on antibody recognition for phenylurea compounds using competitive immunoassay and computational chemistry

“…Computational chemistry not only provided a powerful method to explain the cross-reactivity but also became a useful tool to understand the hapten-antibody recognition mechanism in the absence of antibody physicochemical information. In particular, 2D-QSAR or 3D-QSAR methods, such as CoMSIA and CoMFA Contour, have been used in many studies to investigate the binding activities of polyclonal 14 , 18 , 19 and monoclonal 20 , 21 antibodies against structurally similar compounds. These methods, however, are only suitable for broad-specific antibodies that can recognize a large number of targets and are not suitable for antibodies that only recognize several compounds 22 .…”